Please refer disclaimer Overleaf. C.L.E.D.Medium (with Andrade Indicator) M352 Intended Use: Recommended for isolation and differentiation of urinary pathogens on the basis of lactose fermentation. Composition** Ingredients Gms / Litre Peptone 4.000 HM peptone B # 3.000 Tryptone 4.000 Lactose 10.000 L-Cystine 0.128 Bromothymol blue 0.020 Andrade indicator 0.100 Agar 15.000 Final pH ( at 25°C) 7.5±0.2 **Formula adjusted, standardized to suit performance parameters # - Equivalent to Beef extract Directions Suspend 36.25 grams in 1000 ml of purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates. Principle And Interpretation Sandys reported a new technique where the swarming of Proteus on an agar medium could be prevented by restricting the electrolyte content in the culture medium (7). Sandys Medium was modified by Mackey and Sandys (5), by replacing mannitol with lactose and sucrose and elevating the concentration of agar and bromothymol blue. The same authors further modified this medium by retaining the lactose (deleting sucrose) and by including L-cystine for promoting the growth of cystine-dependent dwarf coliform colony (6). This later modified medium was designated as C.L.E.D. (Cystine- Lactose- Electrolyte-Deficient) Medium. This medium is recommended for use in urinary bacteriology, promoting the growth of all urinary pathogens. C.L.E.D. Medium is also recommended for dip stick procedures and as dip inoculum transport medium for urine specimens (5, 6, 2). C.L.E.D. Medium was further modified by Bevis (1) by incorporation of Andrades indicator. This medium provides sharper differentiation between lactose-fermenters (LF) and lactose-non-fermenters (NLF) (1). Addition of Andrades indicator enhances the appearance of colony and aids in the identification of microorganisms. At different pH values, the colour of the medium varies from the standard medium, which is well documented by Bevis (1). pH Colour of C.L.E.D. medium 7.4 deep blue 7.0 bluish grey 6.8 pale grey 6.6 pinkish grey 6.4 bright red with whitish tinge 6.0 bright red For better results, the medium should not be incubated for more than 24 hours because if lactose fermenters predominate, the entire medium may turn pink masking the presence of non-lactose fermenters. Inoculate the medium immediately after urine
Transcript
Please refer disclaimer Overleaf.
C.L.E.D.Medium (with Andrade Indicator) M352
Intended Use:Recommended for isolation and differentiation of urinary pathogens on the basis of lactose fermentation.
Composition**Ingredients Gms / LitrePeptone 4.000HM peptone B # 3.000Tryptone 4.000Lactose 10.000L-Cystine 0.128Bromothymol blue 0.020Andrade indicator 0.100Agar 15.000Final pH ( at 25°C) 7.5±0.2**Formula adjusted, standardized to suit performance parameters# - Equivalent to Beef extract
DirectionsSuspend 36.25 grams in 1000 ml of purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Sandys reported a new technique where the swarming of Proteus on an agar medium could be prevented by restricting the electrolyte content in the culture medium (7). Sandys Medium was modified by Mackey and Sandys (5), by replacing mannitol with lactose and sucrose and elevating the concentration of agar and bromothymol blue. The same authors further modified this medium by retaining the lactose (deleting sucrose) and by including L-cystine for promoting the growth of cystine-dependent dwarf coliform colony (6). This later modified medium was designated as C.L.E.D. (Cystine- Lactose- Electrolyte-Deficient) Medium. This medium is recommended for use in urinary bacteriology, promoting the growth of all urinary pathogens. C.L.E.D. Medium is also recommended for dip stick procedures and as dip inoculum transport medium for urine specimens (5, 6, 2).C.L.E.D. Medium was further modified by Bevis (1) by incorporation of Andrades indicator. This medium provides sharper differentiation between lactose-fermenters (LF) and lactose-non-fermenters (NLF) (1). Addition of Andrades indicator enhances the appearance of colony and aids in the identification of microorganisms.
At different pH values, the colour of the medium varies from the standard medium, which is well documented by Bevis (1).
pH Colour of C.L.E.D. medium
7.4 deep blue
7.0 bluish grey
6.8 pale grey
6.6 pinkish grey
6.4 bright red with whitish tinge
6.0 bright red
For better results, the medium should not be incubated for more than 24 hours because if lactose fermenters predominate, theentire medium may turn pink masking the presence of non-lactose fermenters. Inoculate the medium immediately after urine
HiMedia Laboratories Technical Data
Please refer disclaimer Overleaf.
collection. Shigella species may not grow on this medium. Prior initiation of antibiotic therapy, low urine pH (less than 5) etc. may result in low urine count from infected patients.
Type of specimen
Clinical:Urine sample
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,4).
Warning and Precautions
In Vitro diagnostic use. Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations1. This medium is recommended for urine infection. Low urine count may be a result of antibiotic therapy, low pH of urine.
2.Recovery depends on the urine count.3.Inoculate the medium immediately after urine collection.4.Shigella species may not grow on this medium.5. For better results, the medium should not be incubated for more than 24 hours because if lactose fermenters predominate,the entire medium may turn pink masking the presence of non-lactose fermenters.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality ControlAppearanceLight yellow to greyish yellow homogeneous free flowing powderGellingFirm, comparable with 1.5% Agar gelColour and Clarity of prepared mediumGreenish blue clear to slightly opalescent gel forms in Petri platesReactionReaction of 3.62% w/v aqueous solution at 25°C. pH : 7.5±0.2pH7.30-7.70Cultural ResponseCultural characteristics observed after an incubation at 35-37°C for 18-24 hours
Storage and Shelf LifeStore between 10-30°C in a tightly closed container and the prepared medium at 20-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition Seal the container tightly after use. Use before expiry date on the label. Product performance is best if used within stated expiry period.
DisposalUser must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
Reference
7. Sandys, 1960, J. Med. Lab. Technol., 17:224.
5. Mackey and Sandys, 1965, Br. Med. J., 2:1286.6. Mackey and Sandys, 1966, Br. Med. J., 1:1173.
2. Dixson J. M. S. and Clark M. A., 1968, Conc. Med. Assoc. J., 99 (15) 1. Bevis T. D., 1968, J. Med. Lab. Technol., 25:38.
Key : *Corresponding WDCM numbers. #- Formerly known as Enterobacter aerogenes
Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained inthis and other related HiMedia™ publications. The information contained in this publication is based on our research and developmentwork and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes tospecifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use butfor laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should notbe considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.
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